Journal: Cell Reports Methods

Article Title: Sequence enrichment profiles enable target-agnostic antibody generation for a broad range of antigens

doi: 10.1016/j.crmeth.2023.100475

Figure Lengend Snippet:

Article Snippet: CD130 , Sino Biological , Cat# 10974-HCCH.

Techniques: Produced, Recombinant, Sequencing, Software

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. qPCR results from pig articular chondrocytes stimulated with OSM and treated with or without SRC inhibitor (SU6656) for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein expression of pSRC in ATDC5 cells after transfection (72 hours) with the modified and WT gp130 plasmids stimulated with OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. qPCR results from ATDC5 cells treated with SRC inhibitor (SU6656) alone, or transfected with indicated variants of plasmids and stimulated with or without OSM for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. D. Protein expression of gp130 pY814 in untreated fetal (17 weeks) and adult articular chondrocytes. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Transfection, Modification

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of pSRC in pig articular chondrocytes stimulated with IL-6 cytokines. Horizontal lines with bars show the mean ± SD. n=3. B. Schematic of modified amino acids within gp130 812-827 domain. Protein expression of pSRC in Ba/F3 cells after transfection with the modified or WT gp130 plasmids stimulated with IL-6 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. CRISPR gRNA was designed by PNA Bio. Y814 mutant mouse (F814) was generated by the USC transgenic mouse core. Mice genotyping was performed by GeneWiz. Representative images of 4-month-old males shown. n=4. D. Protein expression of gp130 pY814 in wild type (WT) and F814 mouse splenocytes treated with and without OSM and LIF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. E. Distribution of differentially expressed genes in MA plot of wild type (WT) and F814 mouse periarticular stromal cells. MA plots (log2fold change vs log2mean expression) for WT-OSM vs WT and Y814-OSM vs Y814 mouse periarticular stromal cells. Each dot represents a gene. Red dots represent upregulated genes i.e. log2fold-change >1 and p-value<0.05. Blue dots represent downregulated genes i.e. log2fold change < −1 and p-value 0.05. Genes that do not qualify this threshold are indicated by grey dots. F. Levels of protein complex formation between gp130 and pSRC in wild type and F814 mouse splenocytes stimulated with or without OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. G. Protein expression of pSRC in wild type and F814 mouse splenocytes stimulated with or without OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Modification, Transfection, CRISPR, Mutagenesis, Generated, Transgenic Assay

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Protein expression of gp130 pY814 in wild type (WT) and F814 mouse splenocytes treated with and without OSM and LIF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein levels of pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. Computational analysis using GOLD software predicted a high affinity binding of peptide QQpYF to the regulatory site of SRC (c-SRC). c-SRC as visualized by crystallography. The structure of the indicated c-SRC domains is shown in ribbon diagram representation (left) as well as with electrostatic potential (blue, positive charge; red, negative charge; white, neutral) mapped onto the molecular surface (right). Peptide QQpYF is shown in stick representation. D. Transcription of genes was determined via qPCR in human adult OA articular chondrocytes treated with or without peptide QQpYF for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. E. Pig knee cartilage explants were stimulated with or without OSM and treated with or without peptide QQpYF at indicated doses for 72 hours followed by a neoepitope assay. Levels of cleaved ACAN and COL2 neoepitopes in the supernatant were quantified with respect to the wet weight of the explant. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Software, Binding Assay

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein levels of pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques:

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of gp130 pY814 in pig articular chondrocytes treated with or without OSM and R805 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Gross morphology and Masson’s trichrome staining of bearded dragon treated with vehicle control and R805 14 days post-amputation. Dashed lines mark amputation planes. n=10. sc=spinal cord; we=wound epithelium; ve=vertebra. C. Gross morphology of bearded dragon co-treated with vehicle control or R805 and PBS or clodronate liposomes 14 days post-amputation. Dashed lines mark amputation planes. n=10. D. Mouse vehicle-treated or R805-treated post-wound day (PWD) 21 wound sections. Representative images are shown. n=8. E. Hair follicle (n=3) and fiber length (n=8) in R805-treated mouse PWD 14 and untreated control wounds. Horizontal lines with bars show the mean ± SD. F. Re-clustering of clusters annotated as macrophages from skin wounds of vehicle-treated or R805-treated mice PWD 14. Dot plots depict gene expression in each macrophage cluster. The contribution of each sample to each cluster is shown as a stacked bar graph. Dot sizes are proportional to the percentage of cells in each cluster expressing the indicated gene. DCs = dendritic cells. G. Re-clustering of clusters annotated as fibroblasts from skin wounds of vehicle-treated or R805-treated mice PWD 14. Dot plots depict gene expression in each fibroblast cluster. The contribution of each sample to each cluster is shown as a stacked bar graph.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Receptor-competition assay. Adult human articular chondrocytes were treated with or without indicated IL-6 family of cytokines and different doses of R805 for 4 hours. Co-IP measuring protein expression was performed after transfection with gp130 FLAG plasmid (72 hours) followed by western blot with respective receptor antibodies. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Competitive Binding Assay, Co-Immunoprecipitation Assay, Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Receptor-competition assay. Adult human articular chondrocytes were treated with or without indicated IL-6 family of cytokines and different doses of R805 for 24 hours. Co-IP measuring protein expression was performed after transfection with gp130 FLAG plasmid (72 hours) followed by western blot with respective receptor antibodies. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Competitive Binding Assay, Co-Immunoprecipitation Assay, Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of gp130 pY814 in pig articular chondrocytes treated with or without OSM and R805 for 4 hours. Horizontal lines with bars show the mean ± SD. n=3. B. qPCR results from pig articular chondrocytes cells treated with OSM with or without R805 for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. C. Pig cartilage knee explants were incubated with or without OSM for 72 hours with or without R805 at indicated concentrations followed by a neoepitope assay. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Incubation

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of R805 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques:

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Affinity kinetics between hIL-6 mutants and hIL-6R − gp130 assayed by SPR. gp130 was immobilized onto a CM5 chips via standard amine coupling and different concentrations of hIL-6 mutants with a saturated concentration of hIL-6R were pre incubated and the mixture were injected. Affinity kinetics were analyzed by the Langmuir’s 1:1 model fit on seven serial dilutions of hIL-6 mutants from 250 nM and flow speed of 30 μL/min. ( A ) Schematic diagram of binding affinity between hIL-6 mutants and hIL-6R − gp130 by SPR; ( B – K ) representative examples of curve fits for the affinity kinetic analysis of hIL-6 mutants.

Article Snippet: The assay was optimized according to the description provided in reference . monomeric gp130 (Sino Biological, Beijing, China, Cat. 10974-HCCH1) was immobilized on CM5 sensor chips (GE Healthcare) by standard amine coupling method, as recommended by the manufacturer.

Techniques: Concentration Assay, Incubation, Injection, Binding Assay

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Affinity of the interaction between hIL-6/mutants and hIL-6R/gp130 by Biacore SPR.

Article Snippet: The assay was optimized according to the description provided in reference . monomeric gp130 (Sino Biological, Beijing, China, Cat. 10974-HCCH1) was immobilized on CM5 sensor chips (GE Healthcare) by standard amine coupling method, as recommended by the manufacturer.

Techniques: Mutagenesis, Binding Assay

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: The spatial arrangement of amino acid residues hIL-6 R167 and E171 in the hexameric hIL-6 − hIL-6R − gp130 complex (PDB code: 1p9m). ( A ) Close spatial proximity is observed between hIL-6 R167 and E171 and the hIL-6 AB-loop; ( B ) the potential influence of hIL-6 R167 on the interaction between hIL-6 K53 and hIL-6R Q95 is indicated. The hIL-6 molecule is shown in brown, hIL-6R in purple, and gp130 in green.

Article Snippet: The assay was optimized according to the description provided in reference . monomeric gp130 (Sino Biological, Beijing, China, Cat. 10974-HCCH1) was immobilized on CM5 sensor chips (GE Healthcare) by standard amine coupling method, as recommended by the manufacturer.

Techniques:

Journal: Cell Reports Medicine

Article Title: Development of a chimeric cytokine receptor that captures IL-6 and enhances the antitumor response of CAR-T cells

doi: 10.1016/j.xcrm.2024.101526

Figure Lengend Snippet:

Article Snippet: Anti-Human CD130-160Gd , Standard BioTools , Cat3160017C.

Techniques: Purification, Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Staining, Antibody Labeling, Mass Cytometry, Mutagenesis, Software, Imaging

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Achieving highly efficient gene transfer to the bladder by increasing the molecular weight of polymer-based nanoparticles.

doi: 10.1016/j.jconrel.2021.02.007

Figure Lengend Snippet: Fig. 6. GP130 protein downregulation mediated by EGCDNPs and Lipofectamine RNAiMAX loaded with siGP130 in UM-UC-3 and UM-UC-3R cells. (A) GP130 and γ-H2AX protein levels assessed by western blot for UM-UC-3 and UM-UC-3R human bladder cancer cells treated with siGP130 – E25GC45DNPs, siGP130 – Lip ofectamine RNAiMAX, or scrambled siRNA (siSC) – E25GC45DNPs at a dose of 4 μg/well siRNA. (B–E) The relative GP130/γ-H2AX expression levels of UM-UC-3 and UM-UC-3R cells are shown. Protein levels were normalized using beta-actin and the data are shown as mean ± SD of three to four independent experiments. * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001.

Article Snippet: Antibodies used included GP130 (NBP2–15776; Novus Biologicals, USA), γ-H2AX (p-Ser 139; Santa Cruz, USA), beta-actin (13E5; Cell Signaling, USA) and a chemiluminescence system was used to detect the protein signal.

Techniques: Western Blot, Expressing

Journal: The Turkish Journal of Gastroenterology

Article Title: IL6ST: A Novel Therapeutic Target for Managing and Treating Colorectal Cancer Via Ferroptosis

doi: 10.5152/tjg.2024.23353

Figure Lengend Snippet: Upregulated IL6ST in CRC tissues and cells was associated with ferroptosis-related genes. (A) IL6ST expression is upregulated in CRC tissues compared with unpaired controls based on TCGA-COAD database ( P = .000). (B) IL6ST expression is upregulated in CRC tissues compared with paired adjacent controls based on TCGA-COAD database ( P = .000). (C) Higher expression of IL6ST in CRC tissues is found compared to adjacent controls via IHC assay ( P = .0001) (200×). (D) IL6ST is upregulated in SW480, HCT116, HT-29, and SW620 cells than nonmalignant human colonic epithelial cell lines NCM-460. (E) Demonstrated the specificity and sensitivity of IL6ST in predicting CRC. (F) The expression level of IL6ST has a significant association with FTH1, GPX4, NCOA4, PTGS2, SLC7A11, and TFRC expression levels.

Article Snippet: For IHC analysis, 5 µm tissue slides were added with a rabbit polyclonal antibody against IL6ST (1:100, diluted with 2% sheep serum, bs-34036R; Bioss, Boston, MA, USA) and incubated at 4°C overnight, followed by blocking with 5% sheep serum for 1 hour at 25°C.

Techniques: Expressing

Journal: The Turkish Journal of Gastroenterology

Article Title: IL6ST: A Novel Therapeutic Target for Managing and Treating Colorectal Cancer Via Ferroptosis

doi: 10.5152/tjg.2024.23353

Figure Lengend Snippet: Knockdown of IL6ST could decrease viability and increase ferroptosis phenotype and related genes. (A-C) Effective knockdown of IL6ST in SW480 CRC cells is detected using RT-PCR. (B, C) Effective knockdown of IL6ST in SW480 CRC cells is detected using western blot. (D) Knockdown of IL6ST could significantly decrease the viability of SW480 CRC cells compared to nonmalignant human colonic epithelial cell lines NCM-460 and vector controls via CCK-8 assays ( P = .007 and P = .004, respectively). (E) IL6ST knockdown SW480 cells exhibit shrunken and damaged mitochondria (3200×). (F) Knockdown of IL6ST up-regulated the levels of iron in SW480 cells compared to the control group ( P = .007). (G) Knockdown of IL6ST increased the levels of ROS in SW480 cells compared to the control group ( P = .005). (H-L) The expressions of PTGS2, NOX1, and ACSL4 are upregulated at the RNA levels, while FTH1 and GPX4 are downregulated in IL6ST knockdown SW480 cells ( P < .01). (M-R) The expressions of PTGS2, NOX1, and ACSL4 are upregulated at protein levels in IL6ST knockdown SW480 cells, while FTH1 and GPX4 are upregulated in IL6ST overexpression SW620 cells ( P < .01).

Article Snippet: For IHC analysis, 5 µm tissue slides were added with a rabbit polyclonal antibody against IL6ST (1:100, diluted with 2% sheep serum, bs-34036R; Bioss, Boston, MA, USA) and incubated at 4°C overnight, followed by blocking with 5% sheep serum for 1 hour at 25°C.

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Plasmid Preparation, CCK-8 Assay, Control, Over Expression

Journal: The Turkish Journal of Gastroenterology

Article Title: IL6ST: A Novel Therapeutic Target for Managing and Treating Colorectal Cancer Via Ferroptosis

doi: 10.5152/tjg.2024.23353

Figure Lengend Snippet: IL6ST overexpression increased cell viability and decreased ferroptosis phenotype and related genes. (A,B) Effective overexpression of IL6ST in SW620 CRC cells is detected using western blot. (C) Overexpression of IL6ST increased the viability of SW620 CRC cells via CCK-8 assays compared to NCM-460 or vector control. (D) IL6ST overexpression protected mitochondrial integrity (3200×). (E) IL6ST overexpression decreased iron accumulation. (F) IL6ST overexpression down-regulated the production of ROS. (G-K) IL6ST up-regulated the expression of FTH1 and GPX4 and decreased expression of PTGS2, NOX1, and ACSL4 at RNA levels ( P < .01).

Article Snippet: For IHC analysis, 5 µm tissue slides were added with a rabbit polyclonal antibody against IL6ST (1:100, diluted with 2% sheep serum, bs-34036R; Bioss, Boston, MA, USA) and incubated at 4°C overnight, followed by blocking with 5% sheep serum for 1 hour at 25°C.

Techniques: Over Expression, Western Blot, CCK-8 Assay, Plasmid Preparation, Control, Expressing

Journal: The Turkish Journal of Gastroenterology

Article Title: IL6ST: A Novel Therapeutic Target for Managing and Treating Colorectal Cancer Via Ferroptosis

doi: 10.5152/tjg.2024.23353

Figure Lengend Snippet: Correlation between IL6ST and immune cell subtype infiltration. (A) The expression level of IL6ST is positively correlated with the infiltration of T helper cells, macrophages, Tcm cells, Tgd cells, Mast cells, DC cells, Th1 cells, TFH cells, Th2 cells, neutrophils and T cells ( P < .05). (B) The enrichment score of T cells is significantly higher in IL6ST high-expressed group than in the low-expressed group.

Article Snippet: For IHC analysis, 5 µm tissue slides were added with a rabbit polyclonal antibody against IL6ST (1:100, diluted with 2% sheep serum, bs-34036R; Bioss, Boston, MA, USA) and incubated at 4°C overnight, followed by blocking with 5% sheep serum for 1 hour at 25°C.

Techniques: Expressing

Journal: Oncology Reports

Article Title: Inhibition of GP130/STAT3 and EMT by combined bazedoxifene and paclitaxel treatment in ovarian cancer

doi: 10.3892/or.2022.8263

Figure Lengend Snippet: Characterisation of bazedoxifene and anti-IL-6 activity in vitro . (A) For surface plasmon resonance (SPR) analysis, bazedoxifene binding to GP130 was immobilised on a CM5 sensor chip and bazedoxifene was injected into the flow cells. (B) T-200 BIA evaluation software was used to subtract the references and determine the steady-state K D . (C) The IL-6-dependent proliferation of DS-1 cells. DS-1 cells were incubated with various concentrations of IL-6 for 72 h for the CCK-8 assay; 10 ng/ml IL-6 was determined as the optimal concentration for subsequent IL-6 inhibitory assays. (D) Inhibition of IL-6-induced proliferation of DS-1 cells by bazedoxifene. DS-1 cells were incubated with IL-6 (10 ng/ml) in the presence of different concentrations of bazedoxifene (*P<0.05, **P<0.01, ***P<0.001). IL-6, interleukin 6.

Article Snippet: The pH scouting for GP130 (Sino Biological, China, or ANRT, Korea) and IL-6Rα (PeproTech) immobilisation was performed in 10 mM acetate buffer at pH 4.0, 4.5, 5.0, and 5.5.

Techniques: Activity Assay, In Vitro, SPR Assay, Binding Assay, Injection, Software, Incubation, CCK-8 Assay, Concentration Assay, Inhibition

Journal: Oncology Reports

Article Title: Inhibition of GP130/STAT3 and EMT by combined bazedoxifene and paclitaxel treatment in ovarian cancer

doi: 10.3892/or.2022.8263

Figure Lengend Snippet: Bazedoxifene inhibits the expression of phospho-STAT3 and estrogen receptor in ovarian cancer cells. (A) Expression of phospho-STAT3 (p-STAT3) and estrogen receptors (ERα and ERβ) was confirmed in ovarian cancer cell lines. (B) After phosphorylation was induced with IL-6 (100 ng/ml) in A2780 and TOV112D cells that did not express p-STAT3, bazedoxifene was administered at different concentrations, showing that p-STAT3 was inhibited following treatment. (C) The degree of p-STAT3 inhibition was confirmed according to the concentration of bazedoxifene in the ER-negative OVCA433 and ER-positive SKOV3 cells. (D) Suppression of p-STAT3 was confirmed by knockdown of STAT3 expression using siRNA. (E) The inhibitory effect of bazedoxifene on the proliferation of OVCA433 and SKOV3 cells after transfection with siNC or siSTAT3-4 is shown (*P<0.05, **P<0.01, ***P<0.001; NS, not significant). STAT3, signal transducer and activator of transcription 3; NC, negative control.

Article Snippet: The pH scouting for GP130 (Sino Biological, China, or ANRT, Korea) and IL-6Rα (PeproTech) immobilisation was performed in 10 mM acetate buffer at pH 4.0, 4.5, 5.0, and 5.5.

Techniques: Expressing, Inhibition, Concentration Assay, Transfection, Negative Control

Journal: Oncology Reports

Article Title: Inhibition of GP130/STAT3 and EMT by combined bazedoxifene and paclitaxel treatment in ovarian cancer

doi: 10.3892/or.2022.8263

Figure Lengend Snippet: The combination of bazedoxifene and paclitaxel inhibits the expression of phosphorylated (p)-GP130, p-STAT3, p-ERK1/2 and EMT-related proteins in ovarian cancer cells. (A) Combined bazedoxifene and paclitaxel decreased the expression of p-GP130, p-STAT3 and p-ERK 1/2 in ovarian cancer OVCA433 and SKOV3 cells. (B) Effect of combined treatment with bazedoxifene and paclitaxel on N-cadherin, E-cadherin, β-catenin, vimentin, Snail, Twist, and Claudin-1 on EMT marker in OVCA433 and SKOV3 cells. EMT, epithelial-mesenchymal transition

Article Snippet: The pH scouting for GP130 (Sino Biological, China, or ANRT, Korea) and IL-6Rα (PeproTech) immobilisation was performed in 10 mM acetate buffer at pH 4.0, 4.5, 5.0, and 5.5.

Techniques: Expressing, Marker

Journal: Oncology Reports

Article Title: Inhibition of GP130/STAT3 and EMT by combined bazedoxifene and paclitaxel treatment in ovarian cancer

doi: 10.3892/or.2022.8263

Figure Lengend Snippet: Combined bazedoxifene and paclitaxel inhibits ovarian cancer OVCA433 tumour growth in vivo . OVCA433 cells (1×10 7 ) were injected subcutaneously into Balb/c-nude mice with an equal volume of PBS. When tumours reached a volume of 200 mm 3 , bazedoxifene (4 mg/kg) with a vehicle of 0.05% CMC and 5% DMSO was administered by gavage five time a week, and paclitaxel (10 mg/kg) was administered by intraperitoneal (i.p.) injection twice weekly. (A) Tumour volumes were calculated from caliper measurements. (B) After 16 days of treatment, all mice were sacrificed, and the total mass (in mg) of each tumour was determined at autopsy (n=6 mice per treatment group). (C) The tumour masses were excised for comparison among the groups. (D) The mouse body weights were assessed on the days indicated. (E) To assess IL-6 production per serum in mouse blood, the total amount of IL-6 was normalised by the vehicle in the different treatment groups (*P<0.05, **P<0.01, ***P<0.001). (F) Hematoxylin and eosin (H&E) staining results show the anticancer effect of combined bazedoxifene and paclitaxel on ovarian cancer. (G) The phosphorylation levels (p) of GP130, STAT3 and ERK1/2 were determined using western blotting of the harvested tumour tissue. β-actin served as the loading control. (H) The protein levels of N-cadherin, E-cadherin, β-catenin, Vimentin, Snail, Twist and Claudin-1 were determined using western blotting of the harvested tumour tissue. β-actin served as the loading control. STAT3, signal transducer and activator of transcription 3; GP130, glycoprotein 130.

Article Snippet: The pH scouting for GP130 (Sino Biological, China, or ANRT, Korea) and IL-6Rα (PeproTech) immobilisation was performed in 10 mM acetate buffer at pH 4.0, 4.5, 5.0, and 5.5.

Techniques: In Vivo, Injection, Staining, Western Blot